Figure 3: GFP-TDP-43-Q303P attenuated ROS-induced cell death and the formation of cytosolic 130-kDa TDP-43 oligomers.
From: Heat-shock protein dysregulation is associated with functional and pathological TDP-43 aggregation
(a) GFP-TDP-43-Q303P increased the resistance to H2O2-induced cell death. All the data are presented as the mean±s.d. (n=3). *P<0.05, t-test. (b) GFP-TDP-43-Q303P overexpression blocked the H2O2-induced increase in the formation of 130-kDa TDP-43 oligomers. (c) H2O2 increased the levels of 130-kDa TDP-43 and the solubility of TDP-43. (d) H2O2 (0.12 mM) increased the cytosolic levels of 130-kDa TDP-43 and increased the nuclear expression of the soluble 43-kDa monomer during a 24-h recovery period after the oxidative insult. (e) Endogenous cytosolic 130-kDa TDP-43 (arrowhead) levels increased in a dose-dependent manner. Scale bars: 10 μm. (f) Cytosolic TDP-43 levels did not increase in GFP-TDP-43-Q303P-expressing cells during a 24-h recovery period after treatment with 0.2 mM H2O2. Scale bars: 10 μm. (g,h) The H2O2-induced formation of 130-kDa TDP-43 was attenuated by NAC. The statistical analysis of the intensity of the 130-kDa TDP-43 bands is presented in g. All the data are presented as the mean±s.d. (n=3). The western blot is illustrated in h. (i) NAC partially suppressed the H2O2-induced increase in endogenous cytosolic TDP-43. The arrowhead indicates NAC-treated cells. The arrow highlights stressed cells that were not inhibited with NAC. Scale bars: 10 μm. Full-sized immunoblots are included in Supplementary Fig. S8.
